EP0563269A1 - Procede de recuperation de natamycine. - Google Patents
Procede de recuperation de natamycine.Info
- Publication number
- EP0563269A1 EP0563269A1 EP92903216A EP92903216A EP0563269A1 EP 0563269 A1 EP0563269 A1 EP 0563269A1 EP 92903216 A EP92903216 A EP 92903216A EP 92903216 A EP92903216 A EP 92903216A EP 0563269 A1 EP0563269 A1 EP 0563269A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- natamycin
- methanol
- water
- solids
- fermentation broth
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/44—Preparation of O-glycosides, e.g. glucosides
- C12P19/60—Preparation of O-glycosides, e.g. glucosides having an oxygen of the saccharide radical directly bound to a non-saccharide heterocyclic ring or a condensed ring system containing a non-saccharide heterocyclic ring, e.g. coumermycin, novobiocin
- C12P19/62—Preparation of O-glycosides, e.g. glucosides having an oxygen of the saccharide radical directly bound to a non-saccharide heterocyclic ring or a condensed ring system containing a non-saccharide heterocyclic ring, e.g. coumermycin, novobiocin the hetero ring having eight or more ring members and only oxygen as ring hetero atoms, e.g. erythromycin, spiramycin, nystatin
- C12P19/626—Natamycin; Pimaricin; Tennecetin
Definitions
- This invention relates to a process of recovery of natamycin from a fermentation broth containing natamycin by adding methanol and adjusting the pH to 1.0 to 4.5 to dissolve the precipitated natamycin; removing the remaining suspended solids; and raising the pH to 6.0 to 9.0 to precipitate high purity natamycin.
- Natamycin is an antibiotic that has been known for decades (Florey, "Analytical Profiles of Drug Substances", Vol. 10, 1981; Merck Index,
- natamycin has a well-defined specific chemical structure, melting point and ultraviolet spectrum. Its specific physic-chemical properties, particularly its solubility in various liquids, require processing techniques specific to this compound. Prior processes for natamycin have not been generally economically acceptable. Processes for recovery of other compounds having different properties give little insight into the recovery of natamycin.
- Natamycin has been prepared by fermentation such as disclosed in British Patent 846,933 (American Cyanamid) using Streptomvces ⁇ ilvosporeus.
- the natamycin . is recovered by methanol -2- extraction followed by tedious steps of adsorption and elution.
- U.S. Patent 3,378,441 discloses recovering natamycin by salting it out of the fermentation broth, extracting with methanol, removing the solids, and then evaporating the liquid to recover the natamycin.
- U.S. Patent 3,892,850 discloses recovery of pimaricin by extraction with acidified butanol followed by distillation and precipitation. It is noted that the pimaricin identification in this patent is inconsistent with those of natamycin as disclosed in Florey.
- natamycin is mechanically removed from the liquid such as by filtration or centrifugation, then washed with water -3- an dried yielding natamycin of over 80% purity and usually over 90% purity.
- steps 2 and 4 sufficient time is allowed to approach stable solubility conditions, usually 15-45 minutes being sufficient depending on the volume of the broth, solids concentration, vessel configura ⁇ tion and the like. Agitation may be helpful.
- the process of the invention requires that the fermentation product plus the extracting liquid contain at least 2 g/1 natamycin. If the direct fermentation broth contains too little natamycin water must be removed. Since the fermentation product consists essentially of water and natamycin plus biomass solids, water can be removed by any convenient solid-liquid separation technique, such as filtering and/or centrifuging off water. Concentrations of up to 50% solids is achievable this way. If a higher solids content is desired in the recovery feed stream to minimize the required methanol, heat assisted vacuum evaporation may be used. Typically a temperature in the range 30*C to 80*C is used. Pro ⁇ longed use of temperatures above 95*C should be avoided to prevent possible product or biomass degra- dation.
- the fermentation product used as the feed stock in the present process comprises solid suspended natamycin and biomass and water.
- the solids content may be as low as 20% or less, for example, if no water is removed from the fermentation broth. If water must be removed to obtain a fermentation product plus extractant containing at least 2 g/1 natamycin, only the minimum water need be removed. Alternatively, depending on equipment availability, particularly for methanol recovery, it may be desirable to remove substantially all of the water before addition of the methanol extractant.
- the concentration of natamycin can be as low as 2 g/1 natamycin.
- the water content of the feed stream is decreased, higher concentrations of natamycin are desirable because of higher solubility of it in methanol containing lower amounts of water.
- the natamycin content preferably is at least 6 g/1.
- methanol is used as the extraction medium because natamycin has excellent solubility in this inexpensive solvent in comparison to higher alcohols, and because simple techniques for recovery of methanol are well known.
- methanol is used in the range of 20 g to about 150 g natamycin per liter of methanol in the feed stream. Since natamycin tends to precipitate out of the acidified methanol/water extraction medium at high concentrations, lower concentration in the range of 20 g to 120 g natamycin per liter methanol is preferred.
- the water content of the methanol extraction liquid increases greater quantities of methanol are needed to achieve the same recovery. For example, if the feed contains 70% water, about three times as much methanol is required than when starting with a substantially dry feed stream. If the feed contains 40% water, twice as much methanol is needed.
- step 2 the pH is adjusted to 1.0-4.5 by the addition of an acidifying agent at any stage, such as directly to the methanol or simultaneously with or after the methanol addition directly to the fermentation product.
- an acidifying agent such as directly to the methanol or simultaneously with or after the methanol addition directly to the fermentation product.
- Hydrochloric acid is an excellent acidifying agent, although any conventional compatible acidic material is acceptable.
- Typical salts that can be used are NaCl, CaCl2 and others such as disclosed in Penick U.S. Patent 3,378,441.
- step 3 the remaining biomass suspended solids are removed by any convenient means such as filtration or centrifugation. These solids, which still contain a significant quantity of natamycin can be put to other valuable uses.
- any compatible basic material can be used to raise the methanol/water/natamycin liquid pH to 6.0-9.0. This pH modification causes precipitation of high purity natamycin, which is removed and dried.
- Typical useful inexpensive compatible basic materials are sodium and potassium hydroxides.
- water may also be added in step 4 to assist in natamycin precipitation. This may be necessary where the water feed stream concentration is low and natamycin concentration is high. The remaining liquid by-product which contains valuable natamycin, fermentation residues, methanol, inorganic salts and water is sent to methanol recovery. Natamycin of about 80% purity can be recovered in the methanol recovery process.
- Fermentation broth is concentrated to 98% solids (plus a small quantity of nutritive residues) by centrifuging followed by drying at 40°C under vacuum.
- the clarified liquid consisting essentially of 61 g/1 natamycin, methanol and a small quantity of water is raised to pH 7.5 by the addition of potassium hydroxide, and let stand for 30 minutes. A thick white precipitate of natamycin forms. The solids are separated by filtration, washed with water and dried at 40°C under vacuum.
- EXAMPLE 2 This example describes the use of fermentation product concentration solely by filtration.
- Fermentation broth is concentrated by filtration to 50% solids. This feed stock containing about 13 g natamycin is slurried in 500 ml of methanol. Thereafter, the procedure of Example 1 is followed except acetic acid and NaOH are used as the acidic and basic pH adjusting additives.
- EXAMPLE 3 This example describes the use of a low solids content fermentation product feed stock.
- Fermentation broth is concentrated by simple mechanical filtration to 23% solids.
- This fermenta ⁇ tion product containing about 2.9 g natamycin is slurried with 40 ml of methanol and the pH reduced to 2.2 by addition of hydrochloric acid. This slurry is agitated for 30 minutes and then the solids are removed by centrifugation followed by filtration.
- the pH of the clarified liquid containing 38 g/1 natamycin is raised to 7.5 by addition of potas ⁇ sium hydroxide and let stand for 30 minutes. A thick white precipitate forms. The solids are separated out by filtration, washed with water, and dried under vacuum. The dry product is 92% pure natamycin.
Landscapes
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Saccharide Compounds (AREA)
Abstract
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US62358590A | 1990-12-07 | 1990-12-07 | |
US623585 | 1990-12-07 | ||
PCT/US1991/008931 WO1992010580A1 (fr) | 1990-12-07 | 1991-12-05 | Procede de recuperation de natamycine |
Publications (2)
Publication Number | Publication Date |
---|---|
EP0563269A1 true EP0563269A1 (fr) | 1993-10-06 |
EP0563269B1 EP0563269B1 (fr) | 1994-10-12 |
Family
ID=24498637
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP92903216A Expired - Lifetime EP0563269B1 (fr) | 1990-12-07 | 1991-12-05 | Procede de recuperation de natamycine |
Country Status (7)
Country | Link |
---|---|
EP (1) | EP0563269B1 (fr) |
JP (1) | JP2908562B2 (fr) |
AU (1) | AU9157091A (fr) |
BR (1) | BR9107207A (fr) |
CA (1) | CA2097840C (fr) |
DE (1) | DE69104624T2 (fr) |
WO (1) | WO1992010580A1 (fr) |
Families Citing this family (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5591438A (en) * | 1990-12-07 | 1997-01-07 | Bio-Technical Resources L.P. | Natamycin recovery |
WO1995011295A1 (fr) * | 1993-10-22 | 1995-04-27 | Gist-Brocades N.V. | Procede ameliore de recuperation a partir d'un bouillon de fermentation aqueux |
US6884611B2 (en) | 1994-01-19 | 2005-04-26 | Sudzucker Aktiengesellschaft | Preparation of acariogenic sugar substitutes |
WO1995027073A1 (fr) * | 1994-03-31 | 1995-10-12 | Pfizer Inc. | Procede de recuperation de natamycine |
US5942611A (en) * | 1995-01-19 | 1999-08-24 | Cultor Ltd. | Process for natamycin recovery |
KR100512331B1 (ko) * | 2002-12-04 | 2005-09-05 | 경상남도 | 스트렙토미세스 나탈렌시스로부터 나타마이신의 생산방법 |
KR100461577B1 (ko) * | 2002-12-04 | 2004-12-17 | 주식회사 바이오알앤즈 | 나타마이신의 정제방법 |
US7816332B2 (en) | 2003-06-02 | 2010-10-19 | Dsm Ip Assets B.V. | Stable aqueous solution of natamycin fungicide |
US7727966B2 (en) * | 2004-10-28 | 2010-06-01 | Dsm Ip Assets B.V. | Stable needle-shaped crystals of natamycin |
CN101062934B (zh) * | 2007-05-31 | 2010-05-19 | 山东鲁抗医药股份有限公司 | 从发酵技术培养物中提取纳他霉素的方法 |
CN109321617B (zh) * | 2018-10-22 | 2020-10-09 | 淮北师范大学 | 一种利用工农业副产物固态发酵合成那他霉素的方法 |
Family Cites Families (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
FR1188428A (fr) * | 1954-11-04 | 1959-09-22 | Olin Mathieson | Procédé pour la récupération de la nystatine |
US3378441A (en) * | 1963-10-17 | 1968-04-16 | S B Penick & Company | Tetraene antibiotic purification |
US4006222A (en) * | 1975-10-20 | 1977-02-01 | E. R. Squibb & Sons, Inc. | Purification of nystatin |
-
1991
- 1991-12-05 BR BR919107207A patent/BR9107207A/pt not_active Application Discontinuation
- 1991-12-05 AU AU91570/91A patent/AU9157091A/en not_active Abandoned
- 1991-12-05 WO PCT/US1991/008931 patent/WO1992010580A1/fr active IP Right Grant
- 1991-12-05 JP JP50366992A patent/JP2908562B2/ja not_active Expired - Lifetime
- 1991-12-05 EP EP92903216A patent/EP0563269B1/fr not_active Expired - Lifetime
- 1991-12-05 DE DE69104624T patent/DE69104624T2/de not_active Expired - Lifetime
- 1991-12-05 CA CA002097840A patent/CA2097840C/fr not_active Expired - Lifetime
Non-Patent Citations (1)
Title |
---|
See references of WO9210580A1 * |
Also Published As
Publication number | Publication date |
---|---|
CA2097840A1 (fr) | 1992-06-08 |
JPH06504908A (ja) | 1994-06-09 |
DE69104624D1 (de) | 1994-11-17 |
AU9157091A (en) | 1992-07-08 |
JP2908562B2 (ja) | 1999-06-21 |
DE69104624T2 (de) | 1995-05-04 |
EP0563269B1 (fr) | 1994-10-12 |
BR9107207A (pt) | 1994-02-08 |
CA2097840C (fr) | 2002-02-05 |
WO1992010580A1 (fr) | 1992-06-25 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP2006296430A (ja) | ナタマイシンの回収 | |
EP0563269B1 (fr) | Procede de recuperation de natamycine | |
EP0220855B1 (fr) | Procédé de récupération de 4,4'-dihydroxydiphénylsulfone à partir d'un mélange d'isomères | |
AU677890B2 (en) | Process for natamycin recovery | |
US5454875A (en) | Softening and purification of molasses or syrup | |
CN114560783B (zh) | 一种转化液中l-酪氨酸的提取方法 | |
MXPA96004372A (en) | Procedure for the recovery of natamic | |
GB2043052A (en) | The purification of dicarboxylic acids | |
US4696817A (en) | Extraction of teichomycin A2 from whole culture fermentation broth | |
US4621153A (en) | Purification and recovery of amino acids | |
US5001270A (en) | Process for recovering 4,4' dihydroxydiphenyl sulfone from an isomer mixture | |
EP0266117B1 (fr) | Procédé pour obtenir des acides aminés purifiés | |
US5093243A (en) | Process for the separation and recovery of nosiheptide | |
US4010167A (en) | Method for the recovery of zearalenone | |
US5391809A (en) | Method for the production of α-L-aspartyl-L-phenylalanine methyl ester hydrochloride | |
US3526662A (en) | Extraction and purification of tetracycline from fermentation broths | |
JP3157724B2 (ja) | インドールの精製方法 | |
US4137405A (en) | Isolation of antibiotic Cephamycin C | |
JP2001046091A (ja) | キシリトールの製造方法 | |
JPH059149A (ja) | 高純度有機酸の回収法 | |
JPH07309901A (ja) | キサンタンガムの精製法 | |
JPH0489479A (ja) | 光学活性トリプトファンの回収方法 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PUAI | Public reference made under article 153(3) epc to a published international application that has entered the european phase |
Free format text: ORIGINAL CODE: 0009012 |
|
17P | Request for examination filed |
Effective date: 19930528 |
|
AK | Designated contracting states |
Kind code of ref document: A1 Designated state(s): CH DE FR GB IT LI NL |
|
17Q | First examination report despatched |
Effective date: 19940127 |
|
GRAA | (expected) grant |
Free format text: ORIGINAL CODE: 0009210 |
|
AK | Designated contracting states |
Kind code of ref document: B1 Designated state(s): CH DE FR GB IT LI NL |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: LI Effective date: 19941012 Ref country code: CH Effective date: 19941012 |
|
REF | Corresponds to: |
Ref document number: 69104624 Country of ref document: DE Date of ref document: 19941117 |
|
ITF | It: translation for a ep patent filed |
Owner name: ING. C. GREGORJ S.P.A. |
|
REG | Reference to a national code |
Ref country code: CH Ref legal event code: PL |
|
ET | Fr: translation filed | ||
PLBE | No opposition filed within time limit |
Free format text: ORIGINAL CODE: 0009261 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: NO OPPOSITION FILED WITHIN TIME LIMIT |
|
26N | No opposition filed | ||
REG | Reference to a national code |
Ref country code: GB Ref legal event code: IF02 |
|
REG | Reference to a national code |
Ref country code: GB Ref legal event code: 732E |
|
NLS | Nl: assignments of ep-patents |
Owner name: DANISCO CULTOR AMERICA,INC. |
|
REG | Reference to a national code |
Ref country code: FR Ref legal event code: TP |
|
PGFP | Annual fee paid to national office [announced via postgrant information from national office to epo] |
Ref country code: FR Payment date: 20110107 Year of fee payment: 20 |
|
PGFP | Annual fee paid to national office [announced via postgrant information from national office to epo] |
Ref country code: IT Payment date: 20101227 Year of fee payment: 20 Ref country code: NL Payment date: 20101224 Year of fee payment: 20 Ref country code: GB Payment date: 20101229 Year of fee payment: 20 |
|
PGFP | Annual fee paid to national office [announced via postgrant information from national office to epo] |
Ref country code: DE Payment date: 20101229 Year of fee payment: 20 |
|
REG | Reference to a national code |
Ref country code: DE Ref legal event code: R071 Ref document number: 69104624 Country of ref document: DE |
|
REG | Reference to a national code |
Ref country code: DE Ref legal event code: R071 Ref document number: 69104624 Country of ref document: DE |
|
REG | Reference to a national code |
Ref country code: NL Ref legal event code: V4 Effective date: 20111205 |
|
REG | Reference to a national code |
Ref country code: GB Ref legal event code: PE20 Expiry date: 20111204 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: NL Free format text: LAPSE BECAUSE OF EXPIRATION OF PROTECTION Effective date: 20111205 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: GB Free format text: LAPSE BECAUSE OF EXPIRATION OF PROTECTION Effective date: 20111204 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: DE Free format text: LAPSE BECAUSE OF EXPIRATION OF PROTECTION Effective date: 20111206 |